Molds cause asthma and allergic respiratory diseases and are associated with other illnesses such as sinusitis, chronic fatigue syndrome, lethargy, or migraines. Mold occurs in damp or water damaged housing. The relationships between mold exposure and health effects are poorly understood. The aim of this project is to develop a multiplex array for mold biomarkers that will measure 4-7 biomarkers, simultaneously, from molds associated with asthma or that are found in water damaged buildings. The Technical Objectives are i) to develop a fluorescent Multiplex Array for Mold Biomarkers (MAMB) on the xMAP system to measure allergens/antigens from Alternaria, Aspergillus fumigatus, A. versicolor, Stachybotrys and Penicillium spp.;ii) to determine the sensitivity, specificity, reproducibility of the MAMB;and iii) to use the MAMB to measure mold exposure in dust samples from environmental studies of damp housing in the U.S. and Canada. The MAMB will use monoclonal antibody pairs for allergen capture and detection and a standard of purified biomarkers for calibration purposes. Current methods for mold testing, such as mold counts or DNA analyses, do not measure specific biomarkers. The MAMB will apply innovative, state-of-the art array technology to assess exposure to specific mold antigens, some of which are known to cause deleterious immune responses. The array for up to 7 biomarkers is expandable to include other mold antigens, as part of Phase II studies. The MAMB will provide quantiative measures of mold biomarker exposure in a single test and will facilitate large scale epidemiologic studies and population surveys on the health effects of environmental exposure to molds.